ZOOTAXAISSN 1175-5326 (print edition)
ISSN 1175-5334 (online edition)Copyright 2013 Magnolia Press
Zootaxa 3683 (2): 159177 www.mapress.com/zootaxa/ Article
Nyx pholeocola, a new genus and cavernicolous species of tribe
Aedini (Diptera: Culicidae) from southern Thailand based on
morphological and molecular data
RALPH E. HARBACH1, IAN J. KITCHING1, C. LORNA CULVERWELL1,
THERESA M. HOWARD1 & YVONNE-MARIE LINTON2,31Department of Life
Sciences, Natural History Museum (NHM), Cromwell Road, London SW7
5BD, UK. E-mail: [email protected]; [email protected];
[email protected]; [email protected] Reed Army
Institute of Research, Entomology Branch, 503 Robert Grant Avenue,
Silver Spring, MD 20910, USA. E-mail: [email protected] Reed
Biosystematics Unit, Museum Support Center, 4210 Silver Hill Rd,
Suitland MD 20746, USA (formerly of NHM)
Nyx Harbach & Linton, gen. nov., is introduced as a new
mosquito genus of tribe Aedini for a previously unknown
cave-dwelling species, Nyx pholeocola Linton & Harbach, sp.
nov., from southern Thailand. A diagnosis of the genus is pro-vided
that features unique anatomical characters of the adult, pupal and
larval stages of the type species. The affinities of Nyx are
discussed in terms of its position in the phylogeny of Aedini. Nyx
is more closely related to Borichinda and Isoaedesthan to other
genera of tribe Aedini. Salient differences that distinguish these
three genera are contrasted. The male and female genitalia, pupa
and fourth-instar larva of the new species are illustrated. DNA
sequence for the second nuclear in-ternal spacer region (ITS2) and
the 658-bp barcode fragment of the mitochondrial cytochrome c
oxidase I (COI) gene re-veal very low similarity with published
sequences, supporting the unique status of the new species.
Key words: Nyx gen. n., Nyx pholeocola sp. n., COI barcode,
ITS2, mosquito, taxonomy, systematics
Biting adults and immature stages of the new species described
in this paper were discovered in the cave at Wat Tham Phanturat,
Khlong Sok, near Khao Sok National Park in Surat Thani Province of
southern Thailand. Adults reared from these larvae were initially
found to be similar to species of genera Borichinda and Isoaedes
using the keys of Rattanarithikul et al. (2010) for the aedine
fauna of Thailand. Upon closer examination of the adults and their
associated larval and pupal exuviae, it became obvious that the
species was very different from the species of those genera.
Furthermore, comparisons of COI and ITS2 sequences with the
extensive collection of COI barcodes in the Barcode of Life
Database (BOLD; www.boldsystems.org) and GenBank, respectively,
showed that the species does not fall within any currently
recognized genus-level taxon of Aedini, and therefore a new
genus-species combination is proposed and described herein.
Material and methods
This study is based primarily on individually reared adults with
associated larval and pupal exuviae (collection SS74). The
specimens were reared from larvae and pupae collected from a
rimstone pool (20 x 50 cm) deep inside a cave in southern
(peninsular) Thailand (see Type Series). In addition, a few females
were collected landing on the collectors within the cave (SS75).
Pinned link-reared adults of both sexes were examined under
simulated natural light; dissected male and female genitalia and
larval and pupal exuviae were studied with differential
interference contrast optics. Anatomical terminology and
abbreviations used in the descriptions and illustrations follow
Harbach & Knight (1980, 1982).
Accepted by G. Courtney: 11 Jun. 2013; published: 3 Jul. 2013
Licensed under a Creative Commons Attribution License
http://www.boldsystems.orgmailto:[email protected]:[email protected]:[email protected]:[email protected]
DNA extraction was carried out using the commercially available
Qiagen DNeasy Kit (Qiagen Ltd, Sussex, England) following the
manufacturers recommended protocol. A 658-bp fragment of the
mitochondrial cytochrome oxidase subunit I locus (COI)
(corresponding to the barcode fragment of the COI) was amplified
using the universal barcoding primers LCO1490 and HCO2198 (Folmer
et al., 1994) according to the protocol of the Mosquito Barcoding
Initiative, as described by Ruiz et al. (2010). The rDNA ITS2 PCR
was carried out using the primers of Collins & Paskewitz (1996)
and the protocols of Linton et al. (2001). Sequencing reactions
were undertaken in both directions using the Big Dye Terminator Kit
on an ABI3770 automated sequencer (PE Applied BioSystems,
Warrington, England) and the original PCR primers. Sequence
chromatograms were edited in Sequencher v. 4.8 (Genes Codes
Corporation, Ann Arbor, MI, USA). Basic Local Alignment Search Tool
(BLAST) searches and the Identification System (IDS) within the
BOLD database were used to determine homology of the sequences with
published or available sequences. DNA aliquots are retained at -70C
in the Frozen Tissue Biorepository of the Smithsonian Institution
Museum Support Center, Suitland, MD for future reference. Sequence
chromatograms and associated data of the new species sequenced in
this study are freely available under the project code NYX within
the Mosquitoes of the World Campaign in the BOLD database
(www.boldsystems.org). GenBank Accession numbers are as follow:
ITS2 (KC860485KC860486) and COI (KC860482KC860484).
The phylogenetic relationships of the new taxon with other
aedine taxa were examined by including character data for the new
species in the morphological data set of Reinert et al. (2009). The
taxa in this data set comprise 270 species, an outgroup of four
non-aedine species and an ingroup of 266 species of Aedini. The new
species was coded for the 336 morphological characters derived from
eggs (the egg of the new species is unknown), fourth-instar larvae,
pupae, and adult males and females listed by Reinert et al. (2009)
(Appendix), using the protocols described therein. Missing data
were coded as ?, characters that could not be scored due to the
absence of homologous structures were coded as and multistate
characters were treated as unordered. For comparison with the
results of Reinert et al. (2009), the data set was analyzed using
maximum parsimony and implied weighting (with values of the
concavity constant, K, ranging from 5 to 17), implemented in TNT
version 1.1 (Willi Hennig Society Edition) (Goloboff et al., 2008).
Tree searches were conducted using all four New Technology search
options: sectorial search, ratchet, tree drifting and tree fusing.
For the ratchet, upweighting and downweighting probabilities were
set to 5% and the number of replicates to 200. The number of cycles
of tree drifting was set to 50. All other search parameters were
left at their default settings. Analyses were terminated once the
most parsimonious (= fittest) cladogram(s) had been found 10 times.
Under Settings, the General RAM was set to 500 Mb and the maximum
number of trees to be held to 10000. Cladograms were rooted between
Culiseta Felt and the remaining taxa.
Nyx Harbach & Linton, gen. nov.
Type species: Nyx pholeocola Linton & Harbach, sp. nov.
Females. Medium-sized mosquitoes. Head: Eyes narrowly separated
above antennae; vertex with large falcate scales and few broad
scales laterally, ocular and interocular areas with smaller falcate
scales, erect scales on occiput and back of vertex, interocular
setae present; antenna longer than proboscis; maxillary palpus
about 0.2 length of proboscis, with 3 normally developed palpomeres
and a minute vestigial fourth palpomere. Thorax: Scutum with
pattern of narrow pale and dark scales; acrostichal and
dorsocentral setae present; scutellum with very narrow scales;
paratergite narrow, without scales or setae; antepronotum with
scales, postpronotum with few narrow scales; postspiracular setae
and scales present. Wing: Remigium without setae; alula with broad
scales on margin. Legs: Tarsi dark-scaled; both ungues of fore- and
midlegs toothed, hindungues slightly smaller, apparently without
teeth. Abdomen: Tergum I nearly completely covered with scales and
setae, laterotergite with scales. Genitalia: Segment VIII retracted
into segment VII; cercus relatively long, more than twice length of
postgenital lobe; insula without setae; 3 spermathecal
Males. Essentially as the females. Head: Antenna about 0.75
length of proboscis, flagellar whorls long, dense,
HABACH ET AL.160 Zootaxa 3683 (2) 2013 Magnolia Press
directed dorsally and ventrally, flagellomeres 12 and 13
elongate; maxillary palpus short, 0.4 length of proboscis,
comprised of 4 palpomeres, fourth vestigial. Legs: Ungues of fore-
and midlegs enlarged, anterior and posterior unguis of each pair
with a tooth. Genitalia: Tergum IX lobes poorly developed,
separated, with rows of setae; sternum IX with 13 medial setae;
gonocoxite elongate, with lateral scales; gonostylus long, slender
at base, distally flattened, expanded and highly setose; claspette
small, tapered to acute apex with a few setae; aedeagus comprised
of 2 tergally bent lateral plates, each plate with apical teeth;
proctiger strongly developed, paraproct with sharp caudal spine,
cercal setae absent.
Pupae. Cephalothorax: Seta 7-CT longer than seta 6-CT. Abdomen:
Seta 3-III long, single; seta 6-III single, seta 6-VII small,
posterior to seta 9; seta 9-IVVI posterior to seta 8, seta 9-VII
single, stout; paddle oval, external and internal margins
Larvae, fourth-instars. Head: Seta 4-C short, with 3 or 4
branches; setae 5,6,8,13-C single; seta 14-C longer than seta 15-C,
single or double; cervical sclerite present. Thorax: Seta 1-P well
developed, longer than seta 2-P; setae 1,3-P usually double (1,2);
seta 5-P double; seta 5-T small, single. Abdomen: Seta 11-I large,
2,3-branched; seta 2-I,II large, 2,3-branched, inserted
anterolateral to seta 1, seta 2-IIIVI relatively small, single or
double, inserted anterior to seta 1; seta 5-IIVII small, usually
single; seta 7-II single, about half length of seta 6-II; seta
9-IIIVI inserted near and anterior to seta 7; seta 10-IIV inserted
lateral to setae 11,12; seta 13-IIIV single. Segment VIII: Comb
scales spine-like, in single row. Siphon: Acus present, detached;
seta 1-S 2,3-branched, inserted distal to pecten; distal 1 or 2
pecten spines usually slightly more widely spaced. Segment X:
Saddle incomplete, relatively large, extending below lateral
midline, acus not developed, posterior margin with spine-like
spicules; seta 1-X single, shorter than saddle; seta 2-X with
multiple branches; seta 3-X single; ventral brush (seta 4-X) with 4
or 4.5 pairs of setae on grid (with only transverse bars) and 1
Eggs. Unknown.Etymology. Nyx is the Greek goddess of the night a
shadowy figure who stood at or near the beginning of
creation and gave birth to a number of personified gods. Her
sparse appearances in mythology reveal her as a goddess of
exceptional power and beauty. Nyx was rarely and only glimpsed in
the shadows of the world, which prompted the generic name proposed
here. The two-letter abbreviation Nx. is recommended for this
Systematics. The generic status and phylogenetic relationships
of Nyx were assessed objectively by including character data for
Nyx in the data set of Reinert et al. (2009). The characters used
in the analyses and their states observed in Nyx are listed in the
Appendix. Analyses using implied weights and values of K ranging
from 5 to 17 each yielded a single most parsimonious (fittest)
cladogram (MPC). With the most extreme weighting functions, K= 5
and 6, Nyx was placed as the sister group to Isoaedes, though the
placement of this pair with other genera was quite different in the
two MPCs. For K = 713, the pattern of relationships among the
genera of the Stegomyia-group (the clade comprising genera Skusea
to Stegomyia) was consistently recovered and identical to that
shown in figure 1 of Reinert et al. (2009), with the addition of
Nyx, which was placed as a separate branch of the main stem between
Isoaedes and Borichinda (Fig. 1). The branching pattern at the base
of the Stegomyia-group was thus: (Skusea (Indusius, Cancraedes))
(Fredwardsius (Isoaedes (Nyx (Borichinda ((Diceromyia, Ayurakitia)
(Dendroskusea, remainder of the Stegomyia-group)))))). With K = 14
and 15, Isoaedes, Nyx and Borichinda formed a clade with the first
two as sister taxa. The weakest weighting with K = 16 and 17 showed
a similar but inverted pattern to that for K = 713: (Dendroskusea
(Diceromyia, Ayurakitia)) (Borichinda (Nyx (Isoaedes
(Fredwardsius((Skusea (Indusius, Cancraedes)), remainder of the
Stegomyia-group))))). Reinert et al. (2009) found a similar plateau
of stability between K = 7 and K = 10, and restricted their
discussion to a consideration of the MPC found with K = 9. As we
found a similar pattern, although now extended to K = 13, and for
ease of comparison, we also restrict subsequent discussion to the
MPC found using K = 9.
The branch supporting Isoaedes as sister to the rest of the
Stegomyia-group is supported by 15 unambiguously optimized but
homoplastic characters (33: 0, 54: 0, 98: 1, 101: 1, 158: 2, 188:
1, 198: 0, 226: 0, 240: 0, 256: 0, 261: 0,262: 1, 266: 1, 273: 0,
326:0), which include nine (boldface) of 10 that supported the same
branch in the study of Reinert et al. (2009). The branch uniting
Nyx with the remainder of the Stegomyia-group to the exclusion of
Isoaedes is supported by seven unambiguously optimized but
homoplastic characters (7:0, 24:1, 34:1, 63:1, 92:1, 290:1, 317:1),
and the branch supporting the clade comprising Borichinda and the
rest of the Stegomyia-group to the exclusion of Isoaedes and Nyx is
supported by 10 unambiguously optimized but homoplastic characters
(40: 1, 52: 2, 55: 2, 58: 1, 64: 1, 67: 1, 110: 1, 232: 1, 255: 0,
328: 1). The emboldened characters in these two lists are those
that supported the clade comprising Borichinda and the rest of the
Stegomyia-group to the exclusion of
Zootaxa 3683 (2) 2013 Magnolia Press 161A NEW GENUS AND SPECIES
Isoaedes in the study of Reinert et al. (2009). Thus, six of the
original 13 characters that support this clade in Reinert et al.
(2009) are now placed on the branch below Nyx and five on the
branch above Nyx. The inclusion of Nyx rendered the optimization of
the remaining character, 53:1, ambiguous. In other words, the
addition of Nyx has simply resulted in the slotting in of this
genus on the original branch between Isoaedes and Borichinda and
the almost equal division of the characters that supported this
branch on either side of the node leading to Nyx. Nyxitself is on a
branch supported by 12 unambiguously optimized but homoplastic
characters: 36: 1, 37: 0, 65: 1, 143: 2, 157: 2, 159: 0, 161: 0,
182: 3, 221: 1, 282: 1, 304: 2 and 308: 1. These differences
clearly support the recognition of Nyx as a new polythetically
diagnosed genus of tribe Aedini.
FIGURE 1. Placement of Nyx in the phylogeny of tribe Aedini when
character data for Nx. pholeocola (Appendix) are added to the data
set of Reinert et al. (2009) and analyzed using implied weighting
and a concavity constant (K) of 9. Supporting characters are
indicated on the branches. All are homoplastic, so Nyx, like most
generic-level taxa of Aedini, is a polythetic genus that is
diagnosed by a unique combination of characters.
Nyx pholeocola Linton & Harbach, sp. n.
Female (Figs 2, 3). As described for genus, bearing the
following features and specific characters listed in the Appendix.
Dark scaling light to dark brown, pale scaling white. Head: Dorsum
with large semi-erect pale falcate scales and few broad spatulate
scales laterally on one or both sides, relatively few pale erect
scales that become darker (brown) laterally. Antenna length about
2.0 mm; pedicel with small pale spatulate scales and flagellomere 1
with few small dark spatulate scales on mesal surfaces. Proboscis
(except labella) and maxillary palpus (Fig. 2D) dark-scaled;
proboscis length about 1.8 mm, slightly longer than forefemur;
maxillary palpus bare beneath, with relatively few setae on dorsal
and lateral surfaces, length 0.3 mm. Thorax: Integument light
brown, darker dorsally. Scutum (Fig. 2B) with pattern of coarser
pale scales on background of finer dark scales, pale scaling as
follows: acrostichal line bifurcating into lateral prescutellar
lines, lateral patch on scutal fossa continuous with anterior
dorsocentral area and posterior fossal line that joins posterior
dorsocentral line and narrow line on margin of supraalar area;
scutellum with pale falcate scales on mid and lateral lobes; brown
setae on antepronotum and posteriorly on postpronotum, golden to
golden-brown setae on upper proepisternum, postspiracular area,
upper and lower mesokatepisternal areas, prealar area, upper
mesepisternal area and 1 or 2 (usually 2) on lower anterior area of
mesepimeron; pale falcate scales on postpronotum and few scattered
pale falcate scales on postpronotum, pale spatulate scales on upper
proepisternum, upper and lower posterior mesokatepisternum and
upper anterior mesepimeron anteroventral to upper mesepimeral
setae. Wing: Length about 3.0 mm, entirely dark-scaled. Halter:
Integument pale, capitellum darker dorsally with faint pale scales.
Legs: Anterolateral surface of forecoxa with pale spatulate scales
and prominent golden-brown setae, midcoxa with pale spatulate
scales on anterior side of mid-lateral row of prominent setae,
hindcoxa without scales, with posterolateral row of prominent
setae; ventral surface of trochanters with pale scales and setae
distally; femora with distinct apical pale rings (knee spots) and
subdued posteroventral pale stripes extending to or near apex;
forefemur length about 1.7 mm. Abdomen: Terga IIVIII with
relatively narrow straight basal pale bands, often narrower
medially on terga VII and VIII and occasionally
HABACH ET AL.162 Zootaxa 3683 (2) 2013 Magnolia Press
obsolescent medially on tergum VIII; sterna dark-scaled from
segment II posteriorly. Genitalia (Fig. 3): Segment VIII fully
retracted into segment VII (Fig. 2F); tergum VIII broad anteriorly,
index about 0.7, relatively long stout setae on caudal margin,
sparse short setae anterior to these, basolateral seta present,
broad scales intermixed with setae; sternum VIII broadened
laterally at midlength, index about 0.6, median posterior
emargination separating broad lateral lobes, setae on caudal 0.67
progressively longer toward margin of lobes, setae 13-S in
posterolaterally directed lateral line, basolateral seta present;
tergum IX index about 0.6, caudal margin with broad median
concavity separating setose lobes; upper and lower vaginal lips
relatively broad, moderately pigmented; postgenital lobe (PGL)
somewhat heart-shaped, middle of posterior margin slightly concave
and lateral margins broadly convex, ventral length about 0.06 mm,
width at midlength about 0.08 mm; cercus with more or less straight
mesal margin and curved outer margin, broad in proximal half and
progressively narrower to blunt apex, distal area of dorsal and
lateral surfaces with scattered setae, length about 0.16 mm,
greatest width about 0.08 mm, index about 2.0, cercus/dorsal PGL
index about 3.2; spermathecal capsules with few small pores near
Male. Generally smaller but otherwise similar to the female
excepting obvious sexual differences. Head: Antenna with strongly
developed whorls of numerous long setae projecting dorsally and
ventrally, 2 terminal flagellomeres disproportionately long
compared to other flagellomeres; length about 1.7 mm. Proboscis
with indication of false joint 0.6 from base. Maxillary palpus
(Fig. 2C, E) about 0.4 length of proboscis, slender, dark-scaled,
palpomeres 13 without scales ventrally, apex of palpomere 3 with
few short setae, palpomere 4 bud-like. Wing: Generally paler, veins
with fewer scales, tertiary fringe scales absent; length about 2.4
mm. Abdomen: Tergum and sternum VIII with straight posterior
margins; sternum VIII (post-rotation dorsal position) mainly
pale-scaled with incomplete posteromedian dark band; tergum VIII
mainly dark-scaled. Genitalia (Fig. 3): Tergum IX lobes weakly
produced, narrowly joined medially by membrane, small setae extend
laterally along posterior margin from apex of each lobe; sternum IX
about as long as broad, with 13 small centrally located setae;
gonocoxite elongate, more or less cylindrical, mesal membrane
developed to apex, basal dorsomesal lobe slightly produced with
dense cluster of relatively long setae, dorsal surface with short
setae, lateral and ventral surfaces with long setae and spatulate
scales, apicodorsal lobe weakly developed with patch of setae;
gonostylus long, about 0.75 length of gonocoxite, narrow
proximally, dorsoventrally flattened and laterally expanded in
distal 0.67, lateral margins of expanded portion lined with
relatively long setae to apex; gonostylar claw relatively long,
slightly longer than 0.1 length of gonostylus, dorsoventrally
flattened and slightly bent ventrad; claspette relatively slender
and slightly compressed, with 1 stout apical seta and a small
dorsomesal seta at its base; proctiger about 0.25 length of
gonocoxite, paraproct distally tapered and bent tergally, without
apical teeth; cercal sclerite poorly defined, more or less
membranous, fused with paraproct; aedeagus with lateral plates bent
tergally, joined distally, tergolateral margin of each lined with
short teeth to apex; paramere nearly as long as aedeagus,
articulated with apex of basal piece near midlength.
Pupa (Fig. 4A, B). As described for genus, bearing the following
features and specific characters listed in the Appendix; character
and positions of setae as illustrated. Cephalothorax: Moderately
but unevenly pigmented, scutum darker posteriorly. Setae 13,6,11-CT
single; setae 4,5,79,10,12-CT branched. Trumpet: Moderately and
evenly pigmented; length about 0.5 mm, tracheoid weakly developed
on one side of trumpet, length about 0.08 mm, pinna about 0.13 mm,
width at midlength about 0. 08 mm, index about 6.3. Abdomen:
Moderately pigmented, posterior area of tergum I and anterior fold
lines of sterna IIVIII darker; length about 3.2 mm. Seta 1-I
broom-like, with short thick stem and numerous (approximately 70)
thin simple branches; setae 3,6-I long, single; seta 11-I usually
present, single; alveolus of seta 13-I usually present; seta 1-II
multiple branched, 1-IIIVII with fewer branches (usually 2) on
succeeding posterior terga; seta 6-IIVI relatively long, usually
single, sometimes double on segments II and III; seta 10-II usually
absent; seta 9-VIII with 24 long aciculate branches, nearly half
length of paddle. Genital lobe: Moderately pigmented; long in male,
length about 0.3 mm, about 0.8 length of paddle; about 0.1 mm in
female. Paddle: Lightly and evenly pigmented, midrib darker, midrib
distinct to distal area before seta 1-Pa; length about 0.8 mm,
width about 0.6 mm, index 1.3; seta 1-Pa long, single.
Larva, fourth-instar (Fig. 5). As described for genus; bearing
the following features and specific characters listed in the
Appendix; character and positions of setae as illustrated (larva
reconstructed from larval exuviae, relative positions of setae may
differ slightly from those shown). Head: Nearly round in dorsal
view, length and width about 0.8 mm; moderately pigmented, collar
darker. Dorsomentum with 79 teeth on either side of slightly larger
median tooth, teeth nearest median tooth abutted, lateral 2 or 3
teeth separated. Setae 5,6,810-C single. Antenna: Moderately
pigmented, bent mesad, length about 0.3 mm; seta 1-A inserted at
mid-length, setae 2,5-A
Zootaxa 3683 (2) 2013 Magnolia Press 163A NEW GENUS AND SPECIES
very long, 2-A nearly twice length of 5-A (Fig. 5B). Thorax:
Integument hyaline, smooth. Seta 1-P double or triple; setae 3,8-P
single or double; setae 14-P,M and 13-T more strongly developed
(longer) than in Isoaedes, 14-P double, 14-M double or triple, 13-T
with 4 or 5 branches; setae 13-M and 8-T double or triple. Abdomen:
Integument hyaline, smooth. Seta 3-IVII normally single, moderately
long, all longer than seta 4; seta 6-IVI with long, stout,
aciculate branches, 6-VI noticeably shorter than 6-IV; seta
2-I,II,VII anterolateral to seta 1, 2-IIIVI more or less directly
anterior to seta 1; seta 8-IIVI normally single; seta 10-IIVI
approaches length of following segment; seta 13-III,IV single or
double, longer than following segment. Segment VIII: Comb comprised
of 69 spine-like scales in single row, with minute basolateral
denticles. Siphon: Moderately pigmented, slightly swollen at level
of seta 1-S, surface smooth; length about 0.91.0 mm, width at base
about 0.3 mm, index 3.03.3; acus present, very small, detached from
siphon; pecten with 1012 spines with few denticles basally on
ventral margin, distal 1 or 2 spines more widely spaced; seta 1-S
branched, inserted more or less at level of distal pecten spine.
Segment X: Saddle moderately pigmented; dorsal length about 0.30.4
mm; siphon/saddle index 2.33.3. Seta 1-X single, simple, slightly
shorter than saddle; setae of ventral brush with 2 or 3 branches,
mostly 3-branched, precratal seta single, simple. Dorsal and
ventral anal papillae same length, short and relatively stout,
slightly longer than saddle.
Egg. Unknown.DNA sequence. ITS2 and COI fragments were amplified
from two specimens (SS74-19 and SS74-40, see type
series below). The sequences of the ITS2 fragments were
identical. These fragments (primers removed), which also include a
short portion of the flanking 5.8S and 28S genes, were 287 bases
long and 50.87% AT rich (65A, 81T, 74C, 67G). The sequence shares
greatest similarity (94%) with that of Borichinda cavernicola
Rattanarithikul & Harbach (GenBank accession EF370410),
reflecting the cladistic relationship noted above. The COI barcode
fragments (658 bp) exhibited no intraspecific variation and the
haplotype was 68.38% AT rich (184A, 266T, 115C, 93G). Despite the
presence of COI sequence for Bc. cavernicola in GenBank (Harbach et
al., 2007), the COI gene of Nx. pholeocola proved to be most
similar (89%) to that of Stegomyia albopicta (Skuse) (HQ398902),
which emphasizes the need for further phylogenetic investigations
before the true affinities of Nyx are resolved. Future studies
should also include DNA sequences for Isoaedes cavaticus
Etymology. The name pholeocola is a Latin noun (masculine or
feminine) derived from pholeos, meaning hole, cave or den, and
-cola, meaning dweller or inhabitant. It was chosen because the
species was discovered in and is only known to inhabit the cave at
Wat Tham Phanturat in Khlong Sok, near Khao Sok National Park in
southern Thailand (see below). The specific name is feminine in
agreement with the gender of Nyx.
Bionomics. Larvae and pupae of Nx. pholeocola were collected
from a rimstone pool (20 x 50 cm) in Wat Tham Phanturat cave,
Khlong Sok, near Khao Sok National Park in Surat Thani Province in
southern Thailand. The water in the pool was clear and cold, 10 cm
deep and devoid of vegetation and plant matter. The cave harbours
an enormous number of bats that are likely to be the primary source
of food for adult females. Females, however, avidly attacked
collectors inside the cave; thus, Nx. pholeocola may have the
potential to opportunistically transmit zoonotic pathogens between
bats and humans.
Distribution. Nyx pholeocola is only known from the cave at Wat
Tham Phanturat Khlong Sok, near the Khao Sok National Park in the
Surat Thani Province of southern Thailand (Fig. 4C).
Type series. Eighty-six specimens: 21 , 2 genitalia, 11 , 4
genitalia, 16 fourth-instar larval exuviae [Le] and 35 pupal
exuviae [Pe]. Holotype, (SS74-24), with LePe on microscope slide,
THAILAND: Surat Thani Province, Khlong Sok (near Khao Sok National
Park), Wat Tham Phanturat cave (08 53' 59.6" N, 98 31' 31.2" E),
shallow rimstone pool (20 x 50cm), 23.i.2008 (Y.-M. Linton et al.).
Paratypes (same data as holotype except specimens of collection
SS75 captured landing on humans in cave; specimens with dissected
genitalia on microscope slides are indicated with an asterisk*), 3
(SS75-1, -3, -4); 7 Pe (SS74-2, -4, -5 [dissected head on
microscope slide], -6*, -8, -10, -13*); 11 LePe (SS74-23, -25 -35,
-36, -37, -38, -39, -40 [ used for DNA extraction; COI (KC860482,
body; KC860484, leg only), ITS2 (KC860486)] -41, -42, -46 [LePe of
-35 to -46 retained in 80% ethanol]); 7Pe (SS74-3*, -7*, -17, -19 [
used for DNA extraction: COI (KC860483), ITS2 (KC860485)], -20*,
-22*, -31); 3 LePe (SS74-9 [dissected head on microscope slide],
-26, -43 [LePe retained in 80% ethanol]); 7 Pe (SS74-11, -14, -15,
-27, -28, -29, -30). All specimens are deposited in the Natural
History Museum, London.
HABACH ET AL.164 Zootaxa 3683 (2) 2013 Magnolia Press
FIGURE 2. Anatomical elements of adult females and males of Nyx
pholeocola. A, Thorax (left side) of a female. B, Mesonotum
(dorsal) of a female. C, Head (left side) of a male showing the
short maxillary palpi. D, Maxillary palpi (dorsal) of a female. E,
Left maxillary palpus (dorsal) of a male. F, Terminal abdominal
segments (dorsal) of a female (segment VIII is fully retracted into
segment VII). Ce = cerci; Pl = palpifer; MPlp = maxillary palpus;
13 = palpomeres; VVII = abdominal segments.
Zootaxa 3683 (2) 2013 Magnolia Press 165A NEW GENUS AND SPECIES
FIGURE 3. Genital structures of Nyx pholeocola (aspects as
indicated). AI, Male genitalia: A, with right gonocoxopodite
removed; B,C, gonocoxopodite; D, tergum IX; E,F, proctiger; G,
aedeagus with parameres; H, claspettes; I, sternum IX. J,K, Female
genitalia: J, principal parts; K, tergum IX. Ae = aedeagus; BP =
basal piece; Ce = cercus; Cl = claspette; Gc = gonocoxite; GC
gonostylar claw; Gs = gonostylus; I = insula; Par = paramere; PGL =
postgenital lobe; Ppr = paraproct; X-Te = tergum X. Scales bars =
0.1 mm (AC,J) and 0.05 mm (DI,K).
HABACH ET AL.166 Zootaxa 3683 (2) 2013 Magnolia Press
FIGURE 4. A,B, Pupa of Nyx pholeocola. A, Left side of
cephalothorax, dorsal to right. B, Dorsal (left) and ventral
(right) aspects of metathorax and abdomen. C, Outline map of
Thailand and Surat Thani Province showing the location of the cave
at Wat Tham Panthurat. CT = cephalothorax; GL = genital lobe; Pa =
paddle; T = trumpet; IVIII, X = abdominal segments IVIII and X;
011, 14 = setal numbers for specified areas, e.g. seta 1II.
Zootaxa 3683 (2) 2013 Magnolia Press 167A NEW GENUS AND SPECIES
FIGURE 5. Fourth-instar larva of Nyx pholeocola (reconstructed
from larval exuviae; positions of some setae may differ slightly
from those illustrated). A, Head, dorsal (left) and ventral (right)
aspects of left side. B, Apex of antenna with setae 26-A. C, Thorax
and abdominal segments IVI, dorsal (left) and ventral (right)
aspects of left side. D, Abdominal segments VIIX, left side. A =
antenna; C = cranium; CS = comb scale; Dm = dorsomentum; M =
mesothorax; P = prothorax; PS = pecten spine; S = siphon; Sa =
saddle; T = metathorax; IVIII, X = abdominal segments IVIII and X;
015, 18 = setal numbers for specified areas, e.g. seta 5-C.
HABACH ET AL.168 Zootaxa 3683 (2) 2013 Magnolia Press
Between 2005 and 2010, Rattanarithikul and her colleagues
published a series of six taxonomic reviews featuring
identification keys that recognized 459 mosquito species
(classified in 53 genera and 40 subgenera) in Thailand. In the
sixth and final publication, Rattanarithikul et al. (2010)
concluded that undescribed species almost certainly will be found
in the future. The discovery of Nyx pholeocola validates this
prediction and brings the total of formally named species of
Culicidae that are known to occur in Thailand to 460 (now
classified in 54 genera).
Nyx is a distinct element of tribe Aedini. The genus and its
sole species can be distinguished from the other genera of the
tribe in Southeast Asia by incorporating the following information
into the keys developed by Rattanarithikul et al. (2010).
Nyx pholeocola keys readily to Plate 12 in the key to adult
females. This couplet distinguishes Isoaedes and Borichinda.
Disregarding character 1, Nyx shares character 2 with Borichinda
(pattern of scutal pale scaling) and character 3 with Isoaedes
(scutellum with narrow scales on midlobe). Character 4 pertains to
postspiracular and subspiracular scales, which are absent in
Isoaedes and present in Borichinda. Nyx differs in having
postspiracular scales but no subspiracular scales.
Nyx pholeocola traces to Borichinda (Bc. cavernicola) at the
bottom of Plate 13 in the key to fourth-instar larvae, with slight
non-conformity of characters in couplets on Plates 9, 11 and 12.
Nyx exhibits the alternative character at the top (right) of Plate
9 and the first alternative (top left) of Plate 11, except that
seta 4-C is not inserted as far anterior to seta 7-C as indicated;
also on Plate 11, seta 4-C does not have many branches as in the
noted exception, Danielsia albotaeniata Leicester. Nyx exhibits two
of the three characters at the bottom (right) of Plate 12, but
disagrees with character 3 in having some unevenly spaced pecten
spines as in the noted exception, Petermattinglyius whartoni
(Mattingly). The characters that identify Borichinda on Plate 13 do
not distinguish Bc. cavernicola from Nx. pholeocola. Characters
that distinguish larvae of Nyx from those of Borichinda are listed
in Table 1, especially the non-stellate condition of setae 1,3-P,
5-T, 2-IVI, 11-I, 5-IIVII, 7-II, 13-IIIV and the precratal seta,
all of which are multiple-branched stellate setae in Bc.
TABLE 1. Salient anatomical differences that distinguish the
adults, pupae and fourth-instar larvae of Nyx, Borichindaand
Isoaedes. Characters that distinguish Nyx from Borichinda are
indicated with an asterisk (*); those that distinguish Nyx from
Isoaedes are indicated with a cross ().
Character Nyx Borichinda Isoaedes
Compound eyes Narrowly separated Narrowly separated
Maxillary palpus, length (males)*
0.3 length of proboscis Slightly shorter than proboscis
0.84 0.92 length of proboscis
Palpomeres (males)* Four, 4th vestigial Five Five
Scutum, pale scaling Present Present Absent
Scutellum, scales on midlobe* Narrow Broad Narrow
Postspiracular scales Present Present Absent
Subspiracular scales Absent Present Absent
Remigial setae* Absent Absent Present
Foreungues (males)* Both toothed One toothed Both toothed
Sternum IX (males) Long Long Shorter
Sternum IX setae Present Present Absent
Gonostylus* Long, enlarged distally Long, cylindrical Long,
Claspette* Apex narrow with few setae Apex expanded with
Apex narrow with few setae
......continued on the next page
Zootaxa 3683 (2) 2013 Magnolia Press 169A NEW GENUS AND SPECIES
Fieldwork during which Nyx pholeocola was discovered was funded
by Royal Caribbean Cruises (The ScholarShip LLC) and Friends of the
Natural History Museum through donations to support the efforts of
the Mosquito Barcoding Initiative (MBI,
http://mosquitobarcode.org/). We are especially grateful to
Ravinder S. Bhatia (Vice President, The ScholarShip) for his
scientific and logistical support, Mark Isenstadt (Director, Vector
Solutions, Bangkok, Thailand) for facilitating field collections,
and Rudy Meiswinkel, Magdalena Zarowiecki and Dean Smallwood for
their assistance in the field. This manuscript was prepared in part
while YML held a National Research Council Research Associateship
Award at the Walter Reed Army Institute of Research. The opinions
and assertions contained herein are those of the authors and are
not to be construed as official or reflecting the views of the US
Department of the Army or the US Department of Defense.
TABLE 1. (Continued)
Character Nyx Borichinda Isoaedes
Seta 7-CT* Longer than 6-CT As long as 6-CT Longer than 6-CT
Seta 3-III Single Single Multiple branches
Seta 6-III Single or double Single Branched
Seta 6-VII* Posteromesal to seta 9 Anterior to seta 9
Posteromesal to seta 9
Seta 9-IVVI* Posterior to seta 8 Anterior to seta 8 Posterior to
Seta 9-VII* Single Branched Branched
Paddle* Apex slightly produced Apex slightly concave Apex
Seta 4-C* Short, 3,4 branches Short, 711 branches Short, 36
Seta 14-C* Single or double, longer than 15-C
Branched, shorter than 15-C
Single, shorter than 15-C
Seta 1-P* Longer than 2-P Shorter than 2-P Longer than 2-P
Setae 1,3-P* Usually double (2,3) Stellate Single
Seta 5-P Double Double Single
Seta 5-T Small single Large, stellate, multiple branches
Seta 2-I,II* Large, 2,3 branches Large, stellate Small,
Seta 2-IIIVI* Small, single or double, anterior to seta 1
Large, stellate, far anterolateral to seta 1
Small, single, near mesal to seta 1
Seta 11-I* Large, 2,3 branches Large, stellate Small, 1,2
Seta 5-IIVI* Small, usually single Large, stellate Small,
Seta 7-II* Single, short (about half length of 6-II)
Short, stellate Long, 1,2 branches
Seta 9-IIIVI* Near anterior to seta 7 Far anterior to seta 7
Near anterior to seta 7
Seta 10-IIV* Lateral to setae 11,12 Mesal to setae 11,12 Lateral
to setae 11,12
Seta 13-IIIV* Single Stellate Single
Seta 5-VII* Small, single Large, stellate Small, 1,2
Comb scales Spine-like, in row Spine-like, in row Scale-like, in
Pecten spines* Distal 1 or 2 spines more widely spaced
Evenly spaced Distal spine more widely spaced
Seta 1-X Small, single Large, branched Small, single
Precratal setae* Present (1, single) Present (2, stellate)
HABACH ET AL.170 Zootaxa 3683 (2) 2013 Magnolia Press
Collins, F.H. & Paskewitz, S.M. (1996) A review of the use
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(1994) DNA primers for amplification of mitochondrial cytochrome c
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Goloboff, P.A., Carpenter, J.M., Aras, J.S. & Esquivel,
D.R.M. (2008) Weighting against homoplasy improves phylogenetic
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Harbach, R.E. & Knight, K.L. (1980) Taxonomists Glossary of
Mosquito Anatomy. Plexus Publishing, Marlton, New Jersey, 415
Harbach, R.E. & Knight, K.L. (1982) Corrections and
additions to Taxonomists Glossary of Mosquito Anatomy. Mosquito
Systematics (for 1981), 13, 201217.
Harbach, R.E., Rattanarithikul, R, Howard, T.M., Linton, Y.-M.
& Kitching, I.J. (2007) Systematics of a new genus and
cavernicolous species of the mosquito tribe Aedini (Diptera,
Culicidae) from Thailand. Proceedings of the Entomological Society
of Washington, 109, 469488.
Linton, Y.-M., Harbach, R.E., Chang, M.S., Anthony, T.G. &
Matusop, A. (2001) Morphological and molecular identity of
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P., Coleman, R.E. & Richardson, J.H. (2010) Illustrated keys to
the mosquitoes of Thailand VI. Tribe Aedini. Southeast Asian
Journal of Tropical Medicine and Public Health, 41 (Suppl. 1),
Reinert, J.F., Harbach, R.E. & Kitching, I.J. (2009)
Phylogeny and classification of tribe Aedini (Diptera:
Culicidae).Zoological Journal of the Linnean Society, 157, 700794 +
2 online appendices.
Ruiz, F., Linton, Y.-M., Ponsonby, D.J., Conn, J.E., Herrera,
M., Quinones, M.L., Velez, I.D. & Wilkerson, R.C. (2010)
Molecular comparison of topotypic specimens confirms Anopheles
(Nyssorhynchus) dunhami Causey (Diptera: Culicidae) in the
Colombian Amazon. Memorias do Instituto Oswaldo Cruz, 105,
Anatomical characters used in the cladistic analysis. See
Reinert et al. (2009) for character coding and discussions of the
characters. Character states observed in Nyx pholeocola are listed
with the numerical character code of Reinert et al. in
1. Oviposition, eggs laid: unknown (?). 2. Anterior end, shape:
unknown (?). 3. Width at midlength: unknown (?). 4. Outer chorion,
cell pattern: unknown (?).
5. Labiogula, length: < width (0). 6. Antenna, ratio of
length to median length of dorsal apotome (DAp): 0.42 (1). 7.
Antenna, spicules: absent (0). 8. Seta 1-A, ratio of length to
antennal width at point of attachment: 3.1 (1). 9. Seta 1-A,
development: 2-branched (0). 10. Setae 2,3-A, insertion on antenna:
apical or nearly apical (0). 11. Seta 1-C, development: single,
thinner, distal part attenuate (1).12. Seta 4-C, insertion relative
to seta 6-C: posterior (2). 13. Seta 4-C, ratio of length to median
length of DAp: 0.19 (0). 14. Seta 5-C, ratio of length to median
length of DAp: 0.410.81 (2). 15. Seta 5-C, development: single (0).
16. Seta 6-C, insertion relative to seta 7-C: anterior (0). 17.
Seta 6-C, ratio of length to median length of DAp: 0.210.38
Zootaxa 3683 (2) 2013 Magnolia Press 171A NEW GENUS AND SPECIES
18. Seta 6-C, development: single (0). 19. Seta 7-C, insertion
relative to seta 5-C: anterior (0). 20. Seta 7-C, ratio of length
to median length of DAp: 0.210.38 (1). 21. Seta 7-C, development: 3
branches (1). 22. Seta 12-C, insertion relative to seta 13-C: mesal
(0). 23. Seta 13-C, development: single (0). 24. Seta 14-C,
development: branched (1). 25. Seta 19-C: absent (0). 26.
Ventromedian cervical sclerite: present (1).27. Setae 13-P,
insertion: not on common support plate (0). 28. Seta 1-P, length
relative to length of seta 2-P: longer (1). 29. Seta 2-P,
development: single (0). 30. Seta 3-P, length relative to length of
seta 2-P: shorter (0).31. Seta 4-P, length relative to length of
seta 3-P: longer (1). 32. Seta 4-P, development: single (0). 33.
Seta 5-P, length relative to length of seta 6-P: equal (0). 34.
Seta 5-P, development: branched (0). 35. Seta 7-P, development:
single or branched (0,1). 36. Seta 8-P, ratio of length to length
of seta 4-P: 1.8 (1). 37. Seta 8-P, development: 2-branched (0).
38. Seta 13-P: absent (0). 39. Seta 1-M, ratio of length to length
of seta 2-M: 2.5 (0). 40. Seta 1-M, development: single (rarely
2-branched) (0). 41. Seta 4-M, development: single (0). 42. Seta
7-M, length relative to length of seta 5-M: shorter (0). 43. Seta
1-T, development: slender (0). 44. Seta 1-T, insertion: on
integument (0). 45. Seta 2-T, development: single (0). 46. Seta
4-T, development: 2 branches, not stellate (1). 47. Seta 6-T,
development: single (0). 48. Seta 3-I, development: single (0). 49.
Seta 7-I, ratio of length to length of seta 6-I: 0.55 (1). 50. Seta
7-I, development: single (0). 51. Seta 12-I: absent (0). 52. Seta
1-II, development: single or 2 branches, not stellate (0,1). 53.
Seta 2-II, development: branched (1). 54. Seta 3-II, development:
single (0). 55. Seta 5-II, development: 2 branches, not stellate
(1). 56. Seta 6-II, length relative to length of seta 6-III: equal
(1). 57. Seta 6-II, development: branched (1). 58. Seta 7-II,
development compared to seta 7-I: similar (0). 59. Seta 8-II,
development: single (0). 60. Seta 6-III, development: branched
(1).61. Seta 3-V, ratio of length to length of setae 5-V: 1.55 (0).
62. Seta 1-VII, ratio of length to middorsal length of segment X:
0.480.85 (1). 63. Seta 2-VII, insertion on tergum VII: on anterior
0.40 far anterior to seta 1-VII (1). 64. Seta 2-VII, development:
single (0). 65. Seta 3-VII, insertion relative to seta 1-VII: at
same level (1).66. Seta 3-VII, development: single (0). 67. Seta
3-VII, ratio of length to middorsal length of segment X: 0.42 (0).
68. Seta 10-VII, development: single (0). 69. Seta 12-VII,
insertion relative to seta 13-VII: posterior (2).70. Seta 12-VII,
development: single (0). 71. Setae 1,2-VIII, insertion: not on
common setal support plate (0). 72. Seta 1-VIII, ratio of length to
length of seta 2-VIII: 0.500.95 (1). 73. Seta 1-VIII, development:
single (0). 74. Seta 2-VIII, development: single (0).75. Seta
2-VIII, insertion: nearer to seta 1-VIII than to seta 3-VIII (0).
76. Seta 3-VIII, development: aciculate (1). 77. Seta 4-VIII,
development: single (0). 78. Comb, scales: several in one row
HABACH ET AL.172 Zootaxa 3683 (2) 2013 Magnolia Press
79. Comb plate: absent (0).80. Siphon, acus: present (1).81.
Pecten: present (1). 82. Pecten spines, arrangement: distal one
more widely spaced (1).83. Seta 1-S, number: 1 (0). 84. Seta 1a-S,
ratio of length to width of siphon: 0.431.12 (1). 85. Seta 1a-S,
development: branched (1). 86. Seta 1a-S, insertion: distal to
pecten (1). 87. Seta 6-S, length relative to distal width of
siphon: equal (0). 88. Seta 8-S, length relative to distal width of
siphon: shorter (0). 89. Seta 9-S, development: slender, slightly
curved (0). 90. Sclerotization of segment X (saddle): incomplete
ventrally (0).91. Saddle, acus: absent (0). 92. Saddle, moderate to
well-developed spicules on posterior margin: present (1). 93. Seta
1-X, insertion: on saddle (0). 94. Seta 2-X, ratio of length to
length of seta 3-X: 0.70 (rarely 0.80) (1). 95. Seta 2-X,
development: 3-branched (0). 96. Seta 2-X, aciculate: absent (0).
97. Seta 3-X, development: single (0). 98. Ventral brush (seta
4-X), attachment: on grid with transverse bars (1). 99.
Precratal/preboss setae (i.e. two or more setae anterior to grid or
boss): present (1). 100. Seta 4a-X of ventral brush, ratio of
length to length of seta 4c-X: 0.70 (1). 101. Seta 4d-X of ventral
brush, development: 3-branched, not plumose (1).
102. Cephalothorax, clear unpigmented spots: absent (0). 103.
Trumpet, tracheoid area: present (1). 104. Trumpet, tracheoid area,
development: weakly developed, at base (0). 105. Seta 1-CT,
development relative to seta 3-CT: weakly developed, considerably
shorter than seta 3-CT (0).106. Seta 5-CT, ratio of length to
length of seta 4-CT: 1.2 (0). 107. Seta 7-CT, length relative to
length of seta 6-CT: 1.25.0 times as long (1). 108. Seta 11-CT,
development: single (0). 109. Seta 13-CT: absent (0). 110. Seta
3-I, length relative to length of seta 6-I: shorter (0). 111. Seta
3-I, development: single (0). 112. Seta 6-I, length relative to
length of seta 7-I: longer (1).113. Seta 1-II, development: 5
branches (1). 114. Seta 2-II, insertion relative to seta 1-II:
lateral (1). 115. Seta 2-II, length relative to length of seta
1-II: shorter (0). 116. Seta 3-II, insertion relative to seta 2-II:
directly posterior (0). 117. Seta 3-II, development: single (0).
118. Seta 3-II, length relative to length of seta 6-II: equal or
shorter (0). 119. Seta 3-II, thickness relative to seta 1-II:
thicker (1). 120. Seta 5-II, insertion relative to seta 4-II:
lateral (0). 121. Seta 5-II, length relative to length of seta
3-II: shorter (0). 122. Seta 6-II, length relative to length of
seta 7-II: longer (1). 123. Seta 3-III, length relative to length
of seta 5-III: longer (1). 124. Seta 3-III, development: single
(0). 125. Seta 6-III, development: single or branched (0,1). 126.
Seta 2-V, insertion relative to seta 3-V: anterior (0). 127. Seta
5-V, length relative to median length of tergum VI: longer (1).128.
Seta 2-VI, insertion relative to seta 1-VI: lateral (1). 129. Seta
3-VI, insertion relative to seta 1-VI: lateral (1). 130. Seta
6-VII, insertion relative to seta 9-VII: posterior (1). 131. Seta
9-VII, length relative to length of seta 6-VII: longer (1). 132.
Seta 9-VIII, insertion relative to posterolateral corner of
segment: on corner (0). 133. Seta 9-VIII, development: 3 branches
(1).134. Paddle, midrib, development: strong, extending to near
apex of paddle (1).135. Paddle, fringe of hair-like spicules:
absent (0). 136. Paddle, development of apical margin: rounded
Zootaxa 3683 (2) 2013 Magnolia Press 173A NEW GENUS AND SPECIES
137. Seta 1-Pa: present (1). 138. Seta 1-Pa, ratio of length to
length of paddle: 0.33 or 0.400.60 (0,1). 139. Seta 1-Pa,
development: single (0). 140. Seta 2-Pa: absent (0).
Adults (females except where otherwise noted)
141. Head, erect scales: present (1). 142. Head, position erect
scales: on occiput and vertex (1). 143. Head, decumbent scales of
vertex: both broad and narrow (2). 144. Head, ocular line width:
narrow (0). 145. Head, ocular scales: all narrow (0).146. Eyes,
immediately above antennal pedicels: narrowly to moderately
separated (1). 147. Interocular space, scales: present (1). 148.
Interocular space, scales: all narrow (0).149. Interocular space,
setae: present (1). 150. Interocular space, setae: 5 (0). 151.
Antennal pedicel, vestiture on mesal surface: present (1). 152.
Antennal pedicel, mesal surface, vestiture, composition: few
scattered scales (not overlapping and not silvery) and setae
(0). 153. Antennal pedicel, lateral surface, scales: absent
(0).154. Apical two flagellomeres, length compared to length of
other flagellomeres (males): disproportionately longer (0). 155.
Antenna, flagellar whorls, development (males): numerous long
setae, normally directed dorsally and ventrally (2). 156. Maxillary
palpus, pale scales: absent (0).157. Maxillary palpomeres,
development (males): 3, palpomere 4 absent or vestigial (2). 158.
Maxillary palpomeres, position of palpomeres 4 and/or 5 relative to
palpomere 3 (males): nearly straight (2).159. Maxillary palpus,
palpomere 3, ratio of length to length of proboscis (males): 0.14
(0). 160. Maxillary palpus, palpomere 5, ratio of length to length
of palpomere 4 (males): no homologous structure (). 161. Maxillary
palpus, ratio of length to length of proboscis (males): 0.27 (0).
162. Maxillary palpus, setae on palpomeres 3 (distally) and 4
(males): few, short to moderately long (0). 163. Proboscis, length
relative to length of forefemur: longer (1).164. Proboscis, pale
scales: absent (0). 165. Proboscis, pale-scaled band near
midlength: absent (0). 166. Antepronota: widely separated (1). 167.
Antepronotal scales: present (1). 168. Antepronotal scales: all
narrow (0). 169. Anterior acrostichal setae: present (1).170.
Posterior acrostichal setae: present (1). 171. Anterior
dorsocentral setae: present (1). 172. Posterior dorsocentral setae:
present (1). 173. Scutal scales: all narrow (0). 174. Scutum, erect
twisted scales: absent (0). 175. Scutal scales, colour: both pale
and dark (1).176. Combined anterior acrostichal and anterior
dorsocentral areas, large patch of pale scales covering anterior
(0). 177. Anterior acrostichal area, pale-scaled stripe: present
(1). 178. Posterior acrostichal area, pale-scaled stripe: present
(1). 179. Anterior dorsocentral area, pale-scaled stripe: absent
(0). 180. Posterior dorsocentral area, pale-scaled stripe: absent
(0). 181. Scutal fossal scales: sparse (0). 182. Scutal fossal
scales, colour: indefinite arrangement of pale and darker scales
(3). 183. Prescutellar area, median and/or posterior parts, scales:
absent (0). 184. Prescutellar setae: present (1). 185. Prescutellar
setae on each side of thorax: 6 (1). 186. Prescutellar area, pale
scales on outer margin mesal to setae: present (1). 187. Antealar
area, scales on anterior part: present (1). 188. Antealar area,
scales on anterior part, colour: all pale (1). 189. Supraalar area,
pale scales: present (1). 190. Anterior supraalar-posterior
antealar area, transverse patch of pale scales: absent (0). 191.
Scutellum, scales on midlobe: all narrow (0). 192. Scutellum,
scales on lateral lobes: all narrow (0).
HABACH ET AL.174 Zootaxa 3683 (2) 2013 Magnolia Press
193. Paratergal scales: absent (0). 194. Parascutellar scales:
absent (0). 195. Mesopostnotal scales: absent (0). 196.
Mesopostnotal setae: absent (0). 197. Postpronotal scales: present
(1). 198. Postpronotal scales: all narrow (0). 199. Prespiracular
setae: absent (0). 200. Postspiracular setae: present (1). 201.
Postspiracular scales: present (1). 202. Hypostigmal scales: absent
(0). 203. Subspiracular scales: absent (0). 204. Upper
proepisternal setae: 14 and 519 (0,1). 205. Upper proepisternal
scales: present (1). 206. Lower proepisternal scales: absent (0).
207. Upper mesokatepisternal setae: present (1).208.
Mesokatepisternal scales: in two patches (1). 209. Upper prealar
setae: 20 (0). 210. Upper prealar scales: absent (0). 211. Lower
prealar scales: absent (0). 212. Mesepimeral scales: present (1).
213. Mesepimeral scales: in one patch (0). 214. Lower anterior
mesepimeral setae: present (1). 215. Mesepimeral fine setae: absent
(0). 216. Metameron, vestiture: absent (0). 217. Metameron,
vestiture: no homologous structure (). 218. Upper calypter, setae
or hair-like scales numerous, 7 (1).219. Upper calypter, setae or
hair-like scales (males): numerous, 7 (1). 220. Alula, marginal
scales: present (1). 221. Alula, marginal scales: broad (1). 222.
Alula, dorsal broad scales: absent (0). 223. Remigium, dorsal
setae: absent (0). 224. Remigium, insertion of dorsal setae: no
homologous structure (). 225. Remigium, ventral setae: absent (0).
226. Costal scales: all dark (0). 227. Dark pigmentation around
radiomedial crossvein and proximal segment of media 3+4: absent
(0). 228. Vein R2, length relative to length of R2+3: longer (1).
229. Anal vein, point of termination: noticeably distad of this
point (1). 230. Wing, fringe scales, colour: uniform (0). 231.
Wing, dorsal tertiary fringe scales on proximal 0.50: present (1).
232. Wing, dorsal tertiary fringe scales on proximal 0.50 (males):
absent (0). 233. Wing, dorsal tertiary fringe scales, colour:
uniform (0). 234. Anteprocoxal scales: absent (0). 235.
Postprocoxal scales: absent (0).236. Hindcoxa, base relative to
dorsal margin of mesomeron: well below (0).237. Fore-, mid- and
hindfemora, complete subapical, pale-scaled bands: absent (0). 238.
Midfemur, median pale-scaled stripe from base to or near apex on
anterior surface: absent (0). 239. Hindfemur, pale scales dorsally
and/or anteriorly at apex: present (1). 240. Hindtibia, scales,
colour: dark only (0). 241. Hindtarsomere 1, basal pale scales:
absent (0). 242. Hindtarsomere 1, one or more median pale-scaled
bands: absent (0). 243. Hindtarsomere 1, apical pale scales: absent
(0). 244. Hindtarsomere 2, basal pale scales: absent (0). 245.
Hindtarsomere 2, apical pale scales: absent (0). 246. Foreungues,
development: both toothed (2).247. Foreungues, development (males):
both toothed, larger one with one tooth (4).248. Midungues,
development (males): both toothed, larger one with one tooth
(4).249. Hindungues, development: both simple (0). 250. Hindungues,
development (males): both simple (0). 251. Abdominal tergum I,
laterotergite, scales: present (1). 252. Abdominal tergum III,
median dorsobasal pale-scaled area: present (1). 253. Abdominal
tergum III, median dorsoapical pale-scaled area: absent (0).
Zootaxa 3683 (2) 2013 Magnolia Press 175A NEW GENUS AND SPECIES
254. Abdominal terga, lateral setae (males): relatively short to
moderately long (0).255. Abdominal segment VII, cross-section
shape: dorsoventrally flattened (1).
256. Intersegmental membrane between segments VII and VIII:
short to intermediate (0). 257. Tergum VIII, development: entirely
sclerotized, rarely with few small, non-sclerotized areas on
distal, lateral and proximal
areas, without rod-shaped structures laterally on each side
(1).258. Tergum VIII, posterior margin: straight (1). 259. Tergum
VIII, length relative to width: shorter (0). 260. Tergum VIII,
moderately long to long seta(e) on lateral margins of proximal
0.40: absent (0). 261. Tergum VIII, insertion of setae: on distal
0.60 or less (0). 262. Tergum VIII, scales: present, 14 scales (1).
263. Sternum VIII, development: entirely sclerotized (3). 264.
Sternum VIII, posterior margin: median emargination separating
broadly rounded lateral lobes (3). 265. Sternum VIII, seta 2-S,
insertion relative to seta 1-S: noticeably posterior (0). 266.
Sternum VIII, scales: present, 10 scales, often covering much of
surface (1). 267. Tergum IX, width/length ratio: 1.8 (1). 268.
Tergum IX, development: single sclerite, with broad emargination on
posterior margin (0). 269. Tergum IX, setae: present (1). 270.
Tergum IX, insertion of setae: on distal area (0). 271. Postgenital
lobe, posterior margin: emarginate (2). 272. Postgenital lobe,
ratio of ventral width at distal 0.20 to cercus width at midlength:
0.66 (1). 273. Postgenital lobe, ventral index: 0.471.64 (0). 274.
Postgenital lobe, insertion of ventral setae: on distal area (0).
275. Upper vaginal sclerite: present (1).276. Lower vaginal
sclerite: absent (0). 277. Insula, development: tongue-like (0).
278. Insular setae: absent (0).279. Insula, insertion of setae: no
homologous structure (). 280. Cercus index: 2.88 (0). 281. Cercal
scales: absent (0). 282. Cercus, distal part: gently oblique
(1).283. Cercus/dorsal postgenital lobe index: 3.20 (0). 284.
Accessory spermathecae: present (1).285. Accessory spermathecae,
development: two large (1).
286. Tergum IX, posterior margin: two moderately broad to broad
lobes (1). 287. Tergum IX, position of lateral lobes on posterior
margin: widely separated (1).288. Tergum IX, setae: present (1).
289. Tergum IX, setae: all slender (0). 290. Sternum IX, vestiture:
present (1).291. Sternum IX, vestiture: setae (0). 292. Gonocoxite,
dorsomesal apical lobe: absent (0). 293. Gonocoxite, dorsomesal
basal lobe: absent (0). 294. Gonocoxite, scales: present (1). 295.
Gonocoxite, mesal surface: entirely membranous (0). 296.
Gonocoxite, seta(e) on basomesal area of dorsal surface: present
(1). 297. Gonocoxite, setal development on basomesal area of dorsal
surface: all slender (0). 298. Gonocoxite, lateral setae: mostly
long (1). 299. Gonocoxite, few to several short blunt-tipped
spiniforms in a row on mesal area of ventral surface: absent (0).
300. Gonocoxite, row or patch of long moderately broad to broad
scales on mesal area of ventral surface: absent (0). 301.
Gonocoxite, row or patch of long narrow lanceolate setae on mesal
area of ventral surface: absent (0). 302. Gonostylus, attachment to
gonocoxite: apical (0). 303. Gonostylus, proximal part: narrow (0).
304. Gonostylus, median part: noticeably broader than proximal part
(2). 305. Gonostylus, distal part: slightly broader than proximal
part (1). 306. Gonostylus, elongate lobe on lateral surface: absent
(0). 307. Gonostylus, insertion of elongate lobe on lateral
surface: no homologous structure ().
HABACH ET AL.176 Zootaxa 3683 (2) 2013 Magnolia Press
308. Gonostylus, moderately broad to broad lobe on median part
of lateral surface: present (1). 309. Gonostylus, hornlike
projection on distal part of lateral surface: absent (0). 310.
Gonostylus, seta(e) on distal 0.33: present (1). 311. Gonostylus,
scales: absent (0).312. Gonostylar claw(s): present (1). 313.
Gonostylar claw(s), number: one (0). 314. Most proximal gonostylar
claw, insertion on gonostylus: at apex (0). 315. Most proximal
gonostylar claw, ratio of length to length of gonostylus: 0.35
(0).316. Most proximal gonostylar claw, development: moderately
broad spiniform (1). 317. Most proximal gonostylar claw, apex:
bluntly pointed (1). 318. Gonostylus/gonocoxite index: 0.73 (3).
319. Claspette: present (1). 320. Claspette, development: single
basal setose plaque, columnar lobe absent (0). 321. Claspette,
subapical thumb-like projection on columnar stem: no homologous
structure (). 322. Claspette, ratio of columnar stem length to
length of aedeagus: no homologous structure (). 323. Claspette,
vestiture: two simple setae (0).324. Claspette filament, distinct
transverse striations: absent (0). 325. Aedeagus, development:
comprised of two lateral plates (aedeagal sclerites) (1). 326.
Aedeagus, width: widest in distal 0.33 (0). 327. Aedeagal teeth:
present (1). 328. Aedeagal teeth, position: on distal 0.55 (0).
329. Aedeagus, small distal spicules: absent (0). 330.
Opisthophallus and prosophallus: absent (0).331. Opisthophallus,
development: no homologous structure (). 332. Proctiger, sternal
arm: absent (0). 333. Proctiger, cercal setae: absent (0).334.
Proctiger, apical teeth on paraproct: present (1). 335. Paraproct,
subapical small knoblike or thumb-like process: absent (0).
336. Habitat of immature stages: fresh-water rock pools (2).
Zootaxa 3683 (2) 2013 Magnolia Press 177A NEW GENUS AND SPECIES
AbstractIntroductionMaterial and methodsTaxonomyNyx Harbach
& Linton, gen. nov.Nyx pholeocola Linton & Harbach, sp.
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